A Syngeneic Mouse B-Cell Lymphoma Model for Pre-Clinical Evaluation of CD19 CAR T Cells
October 16th, 2018
Here, we present a protocol for the production and pre-clinical testing of murine CD19 CAR T cells by retroviral transduction and utilization as a therapy against established syngeneic A20 B-cell lymphoma in BALB/c mice with or without lymphodepleting pre-conditioning.
This method can help us answer key questions in the CAR T-cell immune therapy field, such as the genetic modification of CAR T-cells and combination with other immune therapies.The main advantage of this technique is that allows comparisons between the Lympho replete and the Lympho depleted settings.This allows us to see the role of endogenous immune cells in both disease progression and their interaction with therapeutic CAR T-cells.The implication of this technique as 10 to therapy on diagnosis of solid tumor because the lessons learned in this model can be directly translated to candy's house with other targets such as CEA.To begin this procedure.First, prepare all needed media and cells as outlined in the text protocol.On day one at complete DMEM to 15 square centimeter culture dishes.Seed 7.5 million platinum E-Cells in these dishes and incubate overnight at 37
Overview
This article presents a protocol for producing murine CD19 CAR T cells through retroviral transduction. It explores their application as a therapy against established syngeneic A20 B-cell lymphoma in BALB/c mice, with or without lymphodepleting pre-conditioning.
Key Study Components
Area of Science
- Immunotherapy
- Oncology
- Cellular Biology
Background
- CAR T-cell therapy is a promising approach in cancer treatment.
- Understanding the interaction between CAR T-cells and endogenous immune cells is crucial.
- The study focuses on comparing lympho-replete and lympho-depleted settings.
- Insights gained may be applicable to solid tumors and other therapeutic targets.
Purpose of Study
- To develop a protocol for generating CD19 CAR T cells.
- To evaluate the efficacy of these cells in treating lymphoma.
- To investigate the role of the immune environment in therapy outcomes.
Methods Used
- Preparation of media and cells as per the outlined protocol.
- Seeding of platinum E-Cells in culture dishes.
- Incubation of cells under controlled conditions.
- Assessment of CAR T-cell functionality in different immune settings.
Main Results
- Successful production of murine CD19 CAR T cells.
- Demonstrated differences in therapeutic efficacy based on immune conditioning.
- Insights into the role of endogenous immune cells in therapy response.
- Potential implications for broader applications in oncology.
Conclusions
- The protocol provides a valuable tool for CAR T-cell research.
- Findings contribute to understanding CAR T-cell interactions in vivo.
- Future studies may expand on these findings to improve cancer therapies.
Frequently Asked Questions
Chapters in this video
0:04
Title
0:55
Days 1 & 2: Preparation and Transfection of Plat-E Retroviral Packaging Cell Line
2:11
Day 3: Preparation of Virus-Containing Supernatant for Transduction and Isolation and In Vitro Activation of Mouse Splenic T Cells
4:27
Day 4: Transduction of Mouse T Cells
6:43
Day 5: Round 2 of Transduction
8:33
In Vitro Validation of CAR T Cell Activity
9:53
Results: Pre-Clinical Evaluation of CD19 CAR T Cells
11:32
Conclusion
